Screening for sucrose phosphorylase in exopolysaccharide producing-lactic acid bacteria reveals SPaseWRS-3(1) in Leuconostoc mesenteroides isolated from sugar containing-beverage "wedang ronde" from Indonesia

Sucrose phosphorylase has been known for its activity to generate some intermediate drug precursors. In spite of the broad acceptor specificity of the enzyme, the difference in enzyme specificity between bacterial species was recognized. A number of lactic acid bacteria strains known as exopolysaccharide producer isolated previously from local foods, beverages and soil samples from Indonesia's biosphere were screened for gene encode sucrose phosphorylase using polymerase chain reaction (PCR). A 1476- bp gene fragment from Leuc mesenteroides MBFWRS-3(1) isolated from sugar containing-beverage from Solo, Central Java, called Wedang Ronde, was successfully cloned encoding 492 amino acid residues of sucrose phosphorylase SPaseWRS-3(1) (AN. HM536929). Its predicted molecular mass was found to be closed to the confirmed one by sodium dodecyl sulfate polyacrylamide gel electrophresis (SDS-PAGE) gel (approximately 57 kDa). Although, it revealed high similarity in amino acid sequences to the existing ones (99% identity to 1355SPase), this is the first report on SPase from Indonesia's biosphere. The recombinant SPaseWRS-3(1) expressed by Escherichia coli BL21 Star^™ showed lower activity as compared to the reference SPase, which is 50%.